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Restriction of human adenovirus replication in Chinese hamster cell lines and their hybrids with human cells
R L Radna1, B Foellmer, L A Feldman
1Department of Biological Sciences, Hunter College of the City University of New York, New York 10021.
Abstract:
We have found that the replication of human adenovirus (Ad2) is restricted in multiple Chinese hamster cell lines including CHO and V79. The major site of restriction involves differential accumulation of late viral proteins as demonstrated by immunofluorescence assay and polyacrylamide gel electrophoresis with and without prior immunoprecipitation. Synthesis of fiber and penton base are markedly reduced, whereas others, such as the 100K polypeptide, are synthesized efficiently. This pattern of restriction is similar to that previously reported for Ad2 infection of several monkey cell lines; however, the restriction is more marked in the Chinese hamster cell lines. The restriction is most likely due to a deficient cellular function since stable cell hybrids between V79 or CHO and human cells are permissive for virus replication. By analysis of a series of hybrids with reduced numbers of human chromosomes, fiber synthesis was correlated with the presence of the short arm of human chromosome 3. More hybrids showed restoration of fiber synthesis than production of progeny virus, suggesting that more than one unlinked function is required for the latter.
Insights
Human adenovirus (Ad2) replication is restricted in Chinese hamster cells due to a deficient cellular function. This restriction, particularly affecting late viral protein synthesis, can be overcome by human chromosome 3.
Area of Science:
- Virology
- Cell Biology
- Genetics
Background:
- Human adenovirus (Ad2) replication is known to be restricted in certain non-human cell lines.
- Chinese hamster cell lines, such as CHO and V79, exhibit a significant restriction to Ad2 replication.
Purpose of the Study:
- To investigate the mechanism and cellular basis of Ad2 replication restriction in Chinese hamster cells.
- To identify cellular factors or chromosomal regions involved in overcoming this restriction.
Main Methods:
- Immunofluorescence assays to detect viral protein accumulation.
- Polyacrylamide gel electrophoresis (PAGE) with and without immunoprecipitation to analyze protein synthesis.
- Analysis of stable cell hybrids between Chinese hamster and human cells with varying human chromosome content.
Main Results:
- Differential accumulation of late viral proteins was observed, with reduced synthesis of fiber and penton base, but efficient synthesis of other proteins.
- The restriction was more pronounced in Chinese hamster cells compared to monkey cell lines.
- Stable cell hybrids were permissive for Ad2 replication, indicating a deficient cellular function.
- Fiber synthesis restoration was correlated with the presence of the short arm of human chromosome 3.
Conclusions:
- A deficient cellular function in Chinese hamster cells restricts human adenovirus (Ad2) replication, primarily affecting late viral protein synthesis.
- The short arm of human chromosome 3 plays a role in restoring fiber protein synthesis.
- Multiple unlinked cellular functions are likely required for complete Ad2 replication, as fiber synthesis restoration did not always lead to progeny virus production.